核苷与核苷酸。204. 含6'α-[N-(氨基烷基)氨甲酰氧基]-碳环胸腺嘧啶核苷的寡脱氧核苷酸的合成及其双链体的热稳定性和核酸酶抗性特性

Nucleosides and nucleotides. 204. Synthesis of oligodeoxynucleotides containing 6'alpha-[N-(Aminoalkyl)carbamoyloxy]-carbocyclic-thymidines and the thermal stability of the duplexes and their nuclease-resistance properties.

作者信息

Ueno Y, Karino N, Matsuda A

机构信息

Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan.

出版信息

Bioconjug Chem. 2000 Nov-Dec;11(6):933-40. doi: 10.1021/bc0000684.

DOI:10.1021/bc0000684

PMID:11087344

Abstract

To construct the nuclease-resistant oligodeoxynucleotides (ODNs) with natural phosphodiester linkages, we synthesized ODNs that contain 6'alpha-[N-(aminoalkyl)carbamoyloxy]-carbocyclic-thymidines (4, 5, and 6). The stability of these ODNs to nuclease hydrolysis was examined by using snake venom phosphodiesterase (3'-exonuclease) and nuclease S1 (endonuclease). It was found that the ODNs containing 4, 5, or 6 were more resistant to both the enzymes than the unmodified ODN. These nuclease-resistant properties are noteworthy, since they have natural phosphodiester linkages. Next, the thermal stabilities of duplexes consisting of these ODNs and either the complementary DNA or RNA were studied by thermal denaturation. The ODNs that contain 4 were found to enhance the thermal stability of the duplexes with the complementary DNA, while those containing 5 or 6 decreased the thermal stability of the ODN-DNA duplexes. On the other hand, all ODNs that contained 4, 5, or 6 decreased the thermal stability of the ODN-RNA duplexes.

摘要

为构建具有天然磷酸二酯键的核酸酶抗性寡脱氧核苷酸（ODN），我们合成了含有6'-α-[N-（氨基烷基）氨基甲酰氧基]-碳环胸腺嘧啶核苷（4、5和6）的ODN。通过使用蛇毒磷酸二酯酶（3'-外切核酸酶）和核酸酶S1（内切核酸酶）检测了这些ODN对核酸酶水解的稳定性。发现含有4、5或6的ODN比未修饰的ODN对这两种酶更具抗性。这些核酸酶抗性特性值得关注，因为它们具有天然磷酸二酯键。接下来，通过热变性研究了由这些ODN与互补DNA或RNA组成的双链体的热稳定性。发现含有4的ODN可增强与互补DNA形成的双链体的热稳定性，而含有5或6的ODN则降低了ODN-DNA双链体的热稳定性。另一方面，所有含有4、5或6的ODN均降低了ODN-RNA双链体的热稳定性。